Campylobacter spp.

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Detection of Campylobacter spp  in Chicken
Fecal Samples by Real Time
PCR

Detection of Campylobacter spp in Chicken Fecal Samples by Real Time PCR

Primers and probes for both Campylobacter spp. and Y. ruckeri were located in the 16S rRNA gene sequence. Sequences were chosen by alignment of 16S rRNA gene sequences from Campylobacter spp., Arcobacter spp., and Yersinia spp. using CLUSTALW Multiple Alignment. Recommended guidelines for designing flu- orogenic probes for 5⬘ nuclease assays were followed (29). Campylobacter prim- ers were designed to capture the four Campylobacter species C. jejuni, C. coli, C. lari, and C. upsaliensis, but not necessarily only these species. Y. ruckeri primers were designed to capture exclusively Y. ruckeri. Candidate primers and probes were tested theoretically by comparison to sequence databases (BLAST Se- quence Comparisons [National Institutes of Health]). The primers selected for detection of Campylobacter spp. were campF2 (5⬘-CACGTGCTACAATGGCA TAT-3 ⬘ ) and campR2 (5 ⬘ -GGCTTCATGCTCTCGAGTT-3 ⬘ ), and the TaqMan probe was campP2 (5 ⬘ -FAM-CAGAGAACAATCCGAACTGGGACA- BHQ1-3) (MWG Biotech AG, Ebersberg, Germany). The primers selected for detection of the internal control were yersF1 (5 ⬘ -GGAGGAAGGGTTAAGTG TTA-3⬘) and yersR1 (5⬘-GAGTTAGCCGGTGCTTCTT-3⬘), and the TaqMan probe was yersP1 (5 ⬘ -Hex-GCGAGTAACGTCAATGTTCAGTGC-BHQ1-3 ⬘ ) (MWG Biotech AG).

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Prevalence and pathogen load of Campylobacter spp., Salmonella enterica and Escherichia coli O157/O145 serogroup in sheep faeces collected at sale yards and in abattoir effluent in Western Australia

Prevalence and pathogen load of Campylobacter spp., Salmonella enterica and Escherichia coli O157/O145 serogroup in sheep faeces collected at sale yards and in abattoir effluent in Western Australia

samples and the DNA was extracted and amplified as described above. Mean detection limits, the coefficient of determination-R squared values and percentage relative standard deviation were calculated. Template copy numbers were converted to numbers of organisms present on the basis that purA (Campylobacter spp.), ompF ( S. enterica ) and the hypothetical protein in E. coli O157/O145 (GenBank accession CP008957) were single copy genes[31-34] and that bacterial genomes are haploid. Therefore, the detected plasmid numbers were equivalent to the numbers

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Prevalence of Campylobacter spp. in poultry meat and meat products imported in Republic of Macedonia

Prevalence of Campylobacter spp. in poultry meat and meat products imported in Republic of Macedonia

vinsko meso i MOM-ot koja iznesuva 81.8% odnosno 84%. Vo govedskoto meso i proizvo- dite od govedsko ~adeno meso, koi bea izbra- ni po slu~aen metod za ispituvawe, ne e ut- vrdeno prisustvo na Campylobacter spp. {to uka`uva na faktot deka vakvite vidovi hra- na ne pretstavuvaat zna~aen izvor na infek- cija odnosno kontaminacija. Spored toa, ne- dovolno termi~ki obrabotenoto `ivinskoto meso pretstavuva najgolem rizik za vkrstena kontaminacija na hranata i za infekcija na lu|eto. Osobeno e visok i procentot na MOM- ot vo koj be{e detektiran Campylobacter spp. Bidej}i rizikot pri konsumacija sve`a hrana od animalno poteklo vo korelacija so poja- vata na humana kampilobakterioza e eviden- ten, potrebno e da se poznavaat i na~inite kako da se namali i eliminira rizikot. Ri- zikot od pojavata na zaboluvawe mo`e da bi- de izbegnat so konzumacija na temelno ter- mi~ki obraboteno crveno meso, pile{ko meso i morska hrana, voda od sigurni izvori, kon- sumacija na pasterizirano mleko. Pritoa e va`no pravilno prakti~no rakuvawe so hra- nata kako vo doma{ni uslovi taka i vo ob- jektite vo koi se podgotvuva hranata.

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Coinfection of Enteric Helicobacterspp  and Campylobacter spp  in Cats

Coinfection of Enteric Helicobacterspp and Campylobacter spp in Cats

as from the liver of a dog with hepatitis (6, 12, 17, 54); “Flex- ispira rappini ” strains, which represent at least 10 Helicobacter taxa, have been isolated from feces of mice, sheep, dogs, and humans and have been associated with abortion in sheep (9). Helicobacter pullorum, first isolated from the feces and liver of chickens, also has been cultured from diarrheic humans (53). Helicobacter canadensis, originally misdiagnosed as H. pul- lorum, has been isolated from Canadian patients with diarrhea (14). Helicobacter cinaedi, isolated from the feces of healthy hamsters, also has been recovered from the inflamed lower bowel and blood of immunocompromised adults and children with diarrhea (11, 24, 57). Recently, mixed infections of Heli- cobacter spp. and Campylobacter spp. have been noted in diar- rheic children residing in developing countries (33). The pur- pose of this study is to describe for the first time coinfection of enteric Helicobacter spp. and Campylobacter spp. in cats and to provide molecular characterization of novel Helicobacter spe- cies in these same animals.

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Development and Evaluation of a Multiplex PCR for the Detection of Campylobacter concisus and Other Campylobacter spp  from Gastroenteritis Cases

Development and Evaluation of a Multiplex PCR for the Detection of Campylobacter concisus and Other Campylobacter spp from Gastroenteritis Cases

Campylobacter spp. are fastidious organisms and require a microaerophilic environment to grow. Most labor- atory procedures are optimized to isolate the more common species: C. jejuni and C. coli. However, the other Campylobacter spp. may have additional growth requirements. For example, C. concisus is an emerging patho- gen which requires hydrogen for growth and is a slow growing organism commonly found in the human oral cavity and was first described in 1981 [8]. It is usually associated with healthy than with diseased periodontal sites and in healthy sites it is usually found in shallow rather than in deeper sites [9]. Past studies have associated this species with gingivitis [10], periodontitis [11], and gastritis since 1989 [12]. More recently, it was isolated from foot ulcers [13], other abscesses from different parts of the body [14] and intestinal biopsies and feacal samples of children with Crohn’s disease and patients with inflammatory bowel diseases (IBD) [15] [16]. An early molecular study reported that C. concisus produced cytotonic-like effects on CHO cells and induced in- tracytoplasmic vacuole formation that is similar to H. pylori [17]. Also a recent study in the United Kingdom showed a significantly higher incidence of C. concisus DNA throughout intestinal biopsies of adults with ulcera- tive colitis compared to those of healthy controls [18]. C. concisus requires a hydrogen enriched atmosphere for growth such as that recommended by the Cape Town Protocol [19]. If optimum growth conditions are not ad- hered to, it may result in a reduced isolation rate [20]. In a study in Japan, metagenomic analysis detected DNA sequences of the Campylobacter spp. genome including C. concisus, whilst the standard culture methods were negative [21]. In a recent study in The Netherlands, the detection frequency (DF) of the emerging pathogen C. concisus was found to be at least similar to the DF of C. jejuni when the species was confirmed by PCR product sequencing [22]. In a more recent study in Chile, a significant difference was reported between conventional culture methods and molecular methods used for the detection of Campylobacter spp. and Arcobacter spp. in faecal samples of patients with diarrhea and of controls. Emerging pathogens like C. concisus and C. ureolyticus were only detected by the molecular method used in the study [23].

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Pet Dogs and Chicken Meat as Reservoirs of Campylobacter spp  in Barbados

Pet Dogs and Chicken Meat as Reservoirs of Campylobacter spp in Barbados

Campylobacter spp. are the second most common pathogen isolated from stools of patients with gastroen- teritis in Barbados. The aim of this study was to identify reservoirs of Campylobacter and the likely source(s) of human infection. Fecal specimens from 596 animals and 311 samples of animal food products were analyzed for the presence of Campylobacter spp. by standard culture techniques. Isolates were characterized by conven- tional phenotypic tests, confirmed by latex agglutination and PCR with genus-specific primers, and identified by the use of species-specific primers. High isolation rates were obtained for chickens (94.2%), pigs (90.5%), dogs (46.9%), cats (37.3%), and wild birds (39.3%). Campylobacter was also recovered from monkeys (17.1%) and sheep (4.2%) but not from cows. Chicken meat was frequently contaminated with Campylobacter (58.4%), but its recovery from other animal food products was rare. Campylobacter jejuni was the most commonly identified species in humans (63.6%), chickens (86.6%), dogs (51.5%), and chicken meat (79.8%). Porcine isolates were predominantly C. coli (98.4%), while cats harbored mainly C. upsaliensis and C. helveticus. Wild birds alone carried urease-positive thermophilic campylobacters. C. jejuni and C. coli isolates from different sources were compared with isolates from humans by randomly amplified polymorphic DNA typing with the primers OPA 11 and HLWL 85. Genotyping revealed similarities between isolates from chicken meat and those from humans and could not distinguish between two clinical isolates and four canine strains. Our results suggest that dogs are significant reservoirs of Campylobacter and contribute to human enteric infections and that chicken meat is a likely vehicle for the transmission of campylobacters to humans.

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Emerging trends of antibiotic resistance and risk factors for Campylobacter spp. in commercially produced turkey flocks.

Emerging trends of antibiotic resistance and risk factors for Campylobacter spp. in commercially produced turkey flocks.

It is also of note that the most commonly isolated Campylobacter strains in the turkey feces were also those most commonly isolated from flies, and in agreement with past studies there was no occurrence of fly-positive flocks without the feces yielding positive results as well (Berndtson, 1996). The most common strains were all multidrug resistant and showed resistance to every antimicrobial they were challenged with (with the exception of erythromycin in C. jejuni which has associated fitness costs) (Bolinger & Kathariou, 2017). Recent whole-genome sequencing of two Campylobacter spp. strains (one C. jejuni, ST-1839, accession numbers for genome/plasmid CP017025 to CP017028 from turkey feces, one C. coli, ST-1067, CP017029 to CP017030) from a fly) has revealed the genetic basis for similar multidrug-resistant phenotypes in turkeys and flies (Miller et al., 2016). Both of these strains harbored plasmids containing tet(O) which confers resistance to tetracycline, as well as kanamycin-resistance genes (Lambert, T., Gerbaud, G., Trieu-Cuot, P., Courvalin, 1985; Liebert et al., 2004; Diane E. Taylor & Chau, 1996; Tenover et al., 1989). Resistance to erythromycin in the C. coli strain was conferred by an A2075G substitution in all three copies of the 23S rRNA gene, and The 86 →Ile in gyrA confer resistance to ciprofloxacin and nalidixic acid (Engberg et al., 2001; Jesse et al., 2006; D. E. Taylor & Courvalin, 1988; Zirnstein et al., 2000).

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Campylobacter spp., Salmonella spp., Verocytotoxic Escherichia coli, and Antibiotic Resistance in Indicator Organisms in Wild Cervids

Campylobacter spp., Salmonella spp., Verocytotoxic Escherichia coli, and Antibiotic Resistance in Indicator Organisms in Wild Cervids

Examination was carried out on the fresh sam- ples (which had not been frozen) from 53 red deer, 82 moose, 38 roe deer and 150 wild rein- deer. These were cultivated directly on Campy- lobacter blood free selective agar (Oxoid CM 739) supplemented with cefoperazone, ampho- tericin B and teicoplanin (Oxoid SR 174), and incubated in a microaerophile atmosphere at 37 °C for 2-3 days. Presumptive Campylobacter colonies were confirmed by phase-contrast mi- croscopy. The different species were identified by phenotypic assays, including growth pattern at 42 °C, catalase production and hippurate hy- drolysis.

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Campylobacter spp., Enterococcus spp., Escherichia coli, Salmonella spp., Yersinia spp., and Cryptosporidium oocysts in semi-domesticated reindeer (Rangifer tarandus tarandus) in Northern Finland and Norway

Campylobacter spp., Enterococcus spp., Escherichia coli, Salmonella spp., Yersinia spp., and Cryptosporidium oocysts in semi-domesticated reindeer (Rangifer tarandus tarandus) in Northern Finland and Norway

With regard to the occurrence of possibly zoonotic patho- gens in reindeer, data is rare. In view of the zoonotic enter- opathogens examined in this study, Campylobacter species, Enterococcus species, Escherichia coli, Salmonella species and Yersinia species are among the most important bacteria which cause severe enteric diseases. In detail, various Campylobacter species (C. coli, C. jejuni subspecies jejuni, C. hyointestinalis, C. lari and C. upsaliensis) were isolated from humans with gastro-enteritis [4,5]. Campylobacter hyointes- tinalis was detected in healthy Finnish reindeer [6]. In a recent study about Campylobacter species and other bacte- ria in wild cervids in Norway, no Campylobacter species were detected in wild reindeer [7]. Enterococcus species are known as pyogenic organisms and can cause hospital acquired infections, but are primarily natural inhabitants of the gastrointestinal tract of humans and animals. Ente- rococcus species in reindeer have not yet been described. The virulence and pathogenesis of the examined bacteria depends on different factors. In E. coli, for instance, the ability to cause severe disease in humans and animals is associated with the occurrence of several virulence factors such as shigatoxins. Therefore, the presence of shigatoxin genes can indicate the virulence of certain strains, also known as shigatoxin-producing E. coli (STEC). In 50 fae- cal samples from wild Norwegian reindeer, 42 were posi- tive for E. coli, but no shigatoxin-producing strains were identified [7]. Like STEC, Campylobacter species, Salmo- nella species and Yersinia species are a problem in meat production with a high infection risk for humans who consume contaminated products. Salmonella species have been found to be associated with mortality in reindeer in Finland and Sweden [8]. Among 153 wild Norwegian reindeer, no positive samples for Salmonella species were found [7]. Out of the former eleven known species of Yers-

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Prevalence of Campylobacter spp. among diarrhoeic HIV-patients in Kaduna, Nigeria

Prevalence of Campylobacter spp. among diarrhoeic HIV-patients in Kaduna, Nigeria

of 20.8% among participants that eat chicken. This high prevalence found in chicken could be due to direct contamination from unhygienic chicken handling during processing, cross contamination during evisceration, unhygienic rinsing of multiple carcasses in the same water and cross contamination of the meat slab by meat handlers with the organisms carried from different farms (Olatoye and Ogunsemoyin, 2016). This is because, the intestinal tract of chickens can harbour large amount of Campylobacter species; during processing, may leak or rupture and the contents transferred to the skin of the carcases (Silva et al., 2011). The bacteria will remain in films on the skin and become entrapped into the crevices and channels which provide a favourable environment for cross contamination (Silva et al., 2011). This was in agreement with Olatoye and Ogunsemoyin, (2016) who reported a higher prevalence of 96% from retail chicken sold in Oyo State. Salihu et al., (2009) also reported a prevalence of 81.9% in Sokoto.

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Frecuency and species of Campylobacter spp  in broiler at three levels of the poultry production chain of Costa Rica

Frecuency and species of Campylobacter spp in broiler at three levels of the poultry production chain of Costa Rica

The signs of disease caused by this agent are: diarrhea (might be bloody), abdominal pain, fever, headache, nausea, and Viñas, 2007, Osali et al., 2012). Barré can occur to some patients who have suffered et al., 2004, Farace et al., 2012, In Costa Rica, 23.42kg of food derived from the poultry chain are consumed annually per capita in (Wright, 2010).The high rate of poultry consumption, combined with inadequate treatment during production, sale, and preparation for consumption implicate a higher risk of infection. Because of this, it’s crucial to know Campylobacter spp in the poultry production chain of Costa Rica. This subject is of high importance nationally, and globally, as the poultry chain in Costa Rica has a high economic relevance in local sales, and in In the past, studies conducted in Costa Rica Peña 1996, Rojas et al., 1996, Castrillo 2010) INTERNATIONAL JOURNAL OF CURRENT RESEARCH

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Regulation of Respiratory Metabolism in Campylobacter spp.

Regulation of Respiratory Metabolism in Campylobacter spp.

210. Vugia, D., Cronquist, A., Hadler, J., Tobin-D'Angelo, M., Blythe, D., Smith, K., Lathrop, S., Morse, D., Cieslak, P., Dunn, J., White, P.L., Guzewich, J.J., Henao, O.L., Hoekstra, R.M., Scallan, E., Angulo, F.J., Griffin, P.M., Tauxe, R.V., Barton Behravesh, C. 2008. Centers for Disease Control and Prevention. Preliminary FoodNet Data on the Incidence of Infection With Pathogens Transmitted Commonly Through Food — 10 States, 2007. Morbidity & Mortality Weekly Report 57:366-370. 211. Walker, R. I., M. B. Caldwell, E. C. Lee, P. Guerry, T. J. Trust, and G. M. Ruiz- Palacios. 1986. Pathophysiology of Campylobacter enteritis. Microbiol Rev 50:81-94. 212. Wang, G., S. Angermuller, and W. Klipp. 1993. Characterization of Rhodobacter capsulatus genes encoding a molybdenum transport system and putative molybdenum-pterin-binding proteins. J Bacteriol 175:3031-3042.

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Molecular characterisation of Salmonella enterica serovar Typhimurium and Campylobacter jejuni faecal carriage by captured rangeland goats

Molecular characterisation of Salmonella enterica serovar Typhimurium and Campylobacter jejuni faecal carriage by captured rangeland goats

confirmed by species specific PCR at the hippuricase (hipO) (344 bp amplicon) gene previously described by Persson and Olsen (2005). PCR products were separated by gel electrophoresis and purified using an in-house filter tip method (Yang et al., 2013). Purified PCR products were sequenced using an ABI Prism Dye Terminator Cycle Sequencing kit (Applied Biosystems) using an annealing temperature of 58 °C. Nucleotide sequences were analysed using Chromas lite version 2.0 (http://www.technelysium.com.au) and aligned with reference sequences from GenBank using Clustal W (http://www.clustalw.genome.jp). The target genes and primers used for the amplification and sequencing of S. enterica and Campylobacter spp. isolates and their length are given (Table 1).

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The epidemiology of campylobacter infection in dogs in the context of the risk of infection to humans

The epidemiology of campylobacter infection in dogs in the context of the risk of infection to humans

Although the (age) graph produced did not indicate a linear relationship, results were skewed by a dog of 192 months of age shedding Campylobacter spp. in every sample tested, and a dog aged 132 months shedding C. jejuni in its last sample (i.e. a late shedder). It is difficult to analyse results based on two dogs but if this trend is representive, and older dogs are at a greater risk of carrying Campylobacter spp. than slightly younger adult dogs, this too is probably related to immunity, which can decrease with age (Blount et al., 2005; Greeley et al., 2001; Greeley et al., 1996; Kaszubowska, 2008). There is also evidence to suggest that effects of stress may be more prominent in older adult animals compared to younger adult animals because NA levels in the plasma are higher, spillover into the intestine is greater, and clearance is poorer in older animals at baseline and during stress (McCarty et al., 1997). Caution must be taken in interpreting the results of this current study because the majority of the dogs in the rescue kennel had their age estimated, and because C. jejuni and C. upsaliensis, which may have different roles, were analysed together.

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Use of Culture, PCR Analysis, and DNA Microarrays for Detection of Campylobacter jejuni and Campylobacter coli from Chicken Feces

Use of Culture, PCR Analysis, and DNA Microarrays for Detection of Campylobacter jejuni and Campylobacter coli from Chicken Feces

Poultry and poultry products are considered important sources of human campylobacteriosis and play important roles in disease transmission (5, 6, 8, 11). In Denmark, a systematic sampling procedure for continued evaluation of the prevalence rates and epidemiology of Campylobacter in poultry has been in place since 1998 (3). At the retail level, 30 to 40% of poultry at slaughter have been reported to be contaminated with Campylobacter (31). Isolation and identification of Campy- lobacter by the conventional culture method are laborious due to the slow growth rate, the lack of phenotypic differences in the bacteria, and often, the failure to identify Campylobacter to the species level by the available culture methods. There is a need for the development of a sensitive, rapid method for Campylobacter detection and identification to the species level. Several PCR assays have successfully been applied to the detection of Campylobacter spp. in water (15, 26), some dairy products (9, 12, 29, 32), and chicken litter (13). The PCR method allows detection not only of viable bacteria but also of noncultivable forms of Campylobacter (12, 32). A multiplex PCR assay suitable for mass screening to detect Campylobacter directly from chicken feces has been developed (1). Agarose gel electrophoresis is often used for the PCR assays. Gel elec-

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Multiplex Real Time PCR for Detection of Campylobacter, Salmonella, and Shigella

Multiplex Real Time PCR for Detection of Campylobacter, Salmonella, and Shigella

In this study, we first standardized a conventional PCR us- ing primers that were previously reported for use against 16S rRNA (Salmonella spp. and Campylobacter spp.) (10, 18, 22) and ipaH (Shigella spp.) (7). Unfortunately, when we put all the primers in a single PCR, we obtained cross-reactions. Conven- tional PCR requires amplification in a thermocycler followed by product separation by gel electrophoresis. The cost, the time delay required to do gel analysis of PCR products, and the inability to analyze large numbers of strains are major imped- iments to this approach. In addition, while ethidium bromide is a relatively inexpensive reagent for DNA gel staining, it has human and environmental safety concerns. For these reasons, real-time fluorescence-based multiplex PCR has become an at- tractive technique. It offers the advantages of being a faster and more robust assay because it does not require post-PCR proce- dures to detect amplification products.

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Prevalence, genotyping and risk factors of thermophilic Campylobacter spreading in organic turkey farms in Germany

Prevalence, genotyping and risk factors of thermophilic Campylobacter spreading in organic turkey farms in Germany

The pathways by which poultry flocks acquire Campylo- bacter are not yet fully understood in detail. The same applies for the formation of the high genetic diversity of Campy- lobacter which was observed in infected poultry flocks of different ages [5–7]. Horizontal transmission is generally considered to be the most significant mode of Campylo- bacter earning by poultry flocks [8–10]. However, the pres- ence of a specific genotype in the environment of the birds does not in itself prove that also the birds are infected [11]. Assumed risk factors and vectors involved in the spreading are beside wild birds and their faeces insects such as darkling beetles and drinking water. Several studies have shown that beetles were only Campylobacter positive when the herd was positive, too [10, 12]. Darkling beetles can play a role in the entry of Campylobacter into a broiler flock [13, 14]. Drinking water can be an important vehicle for Campylobacter spp. transmission to the entire flock [15–17].

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Campylobacterantimicrobial resistance in Peru: a ten year observational study

Campylobacterantimicrobial resistance in Peru: a ten year observational study

Isolates identified as Campylobacter spp. from stool at study sites as part of routine clinical practice were placed into Cary-Blair transport media and stored at 4°C at the study site. Every fifteen days, isolates from each study site were transported to the U.S Naval Medical Re- search Unit-6 (NAMRU-6) laboratory at 4°C for con- firmation of identification, speciation and antimicrobial susceptibility testing. Bacterial isolates received from study sites were re-cultured and re-isolated according to methods described elsewhere [19,20]. Briefly, Campylo- bacter blood-free selective agar media with ECR 0155 supplement (Oxoid, Cambridge, UK) were inoculated and incubated in a microaerophilic environment with a gas mixture of 10% CO2, 5% O2, and 85% N2 in anaer- obic jars at 42°C for 24 to 48 hours. Campylobacter jejuni was differentiated from other Campylobacter spe- cies, including C. coli, by hippurate hydrolysis assay.

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Antibody Responses to CampylobacterInfections Determined by an Enzyme-Linked Immunosorbent Assay: 2-Year  Follow-Up Study of 210 Patients

Antibody Responses to CampylobacterInfections Determined by an Enzyme-Linked Immunosorbent Assay: 2-Year Follow-Up Study of 210 Patients

Together with Salmonella serovars, Campylobacter spp. are the most common bacterial enteric pathogens in developed countries, and Campylobacter jejuni is now the most recognized antecedent cause of Guillain-Barre´ syndrome (15, 16, 21). In Denmark, the incidence of registered Campylobacter infections has increased markedly since 1992 (from 22 cases per 100,000 inhabitants in 1992 to 78 cases per 100,000 in 1999), and a similar emergence of Campylobacter has been observed in oth- er industrialized countries (6). The diagnosis of Campylobacter infections is routinely done by stool culturing on selective me- dium, and C. jejuni and Campylobacter coli account for 94 and 6%, respectively, of Danish human isolates (17). Furthermore, culturing of stools is not a sensitive method for detection of the bacteria in patients treated with antibiotics or in patients with late reactive complications such as arthritis and Guillain-Barre´ syndrome or long-lasting intestinal distress (16). In these cases and for epidemiological studies in general, serodiagnosis is valuable. Antibodies to C. jejuni and C. coli can be detected in several test systems with various sensitivities using a homolo- gous strain or selected reference strains in crude antigen prep- arations.

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Differentiation of Campylobacter coli, Campylobacter jejuni, Campylobacter lari, and Campylobacter upsaliensis by a Multiplex PCR Developed from the Nucleotide Sequence of the Lipid A Gene lpxA

Differentiation of Campylobacter coli, Campylobacter jejuni, Campylobacter lari, and Campylobacter upsaliensis by a Multiplex PCR Developed from the Nucleotide Sequence of the Lipid A Gene lpxA

tions have uncovered numerous point sources for these organ- isms, but in spite of this, there has been no apparent reduction in reported incidences. Molecular assays, such as the multiplex PCR assay developed in this study, should further expand our FIG. 2. Phylogenetic analysis of the lpxA gene of 53 isolates of thermotolerant Campylobacter spp. The lpxA gene from isolates of C. coli, C. jejuni, C. lari, and C. upsaliensis was amplified from genomic DNA by PCR with primers lpxAF0301 and lpxAR0304. Amplicons were sequenced, and the primer regions were removed to facilitate the alignment. In total, 746 bp of C. jejuni, and C. lari nucleotide sequence, 754 bp of C. upsaliensis nucleotide sequence, and 757 bp of C. coli nucleotide sequence were aligned. The lpxA gene of Helicobacter hepaticus (ATCC 51499) was used to root the tree. The scale at the bottom of the figure is a measure of genetic identity. Numbers at the branches indicate the percent bootstrap support for each node. Note that the following isolates were not included in the figure for clarity; the lpxA sequences from these isolates were represented as follows. C. coli RM3232 also represents isolates RM3230 and RM3231 and M275 also represents isolates RM1051, RM1166, RM1505, RM1530, RM1531, RM1533, RM2225, and RM2228. C. jejuni RM3664 also represents isolates RM3665, RM3666, and RM3667; NCTC 11168 also represents ANR0493; RM3668 also represents RM3669 and RM3670; and F38011 also represents isolates RM1221, RM3672, and RM3673. C. lari RM2100 also represents RM2099, RM2808, RM2809, RM2817, RM2820, and RM2826 and RM1890 also represents RM2810 and RM2821. C. upsaliensis: RM1488 also represents RM2094.

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